Physiologically effective substance and process of preparing same



Patented Oct. 21, 1941 UNITED QsTATES PATENT OFFICE l $59,698 j I PHYSIOLOGICALLY EFFECTIVE SUBSTANCE AND PROCESS-0F PREPARING SAME 7 Fritz 'Johannessohiij Mannheim-Feudenhim; and Hans Hatzig'; Mannheim-Waldhof, ,Germany; assignor's to Bare Chemicals Inc., Nepera Park, N. Y., a corporationof NewXo -k r i No Drawing App ication May 6, 1939, SerialNo;

flg i'l-g Jn.e anx-Me .t,,. 93 efs-"cr n (c1. zct apyis) Ou l ve i el tes nhrsiqlqs allmfie tire unds f h clo en a hr to henanthrene series and to pro factureof same.

ces ses for the i It is, W now qj o etskille in arttha b the re me of s ew J91 th ir erivative still containing the side chain with oxidising agents le Q s ti g;ef przer e ly 'QQ pletely the side chain, such as chromic acid or potassium permanganatejone may procure iihy ologicallyvaluable substances ihus, by the dation of the cholesterol} cholestenone, etc., tor example;one obtains" substances-which {possess the activity'of a male sxualj hormone on of a corpus luteum' hormone, .as for instance dehydroandrosterone, androstefidigonef 'fprogesterone and the like.' The substances" ma'y be separated from the oxidation mixtur'e'bylyarious methods, whereby. at. first the VO IatiIE-aIid fiacid degradation products must be removed. ".TheNremoval .ofthe by means of alkaline,;ageiits;,;such(fas laduous alkali hydroxide, wherebygimmost" cultly soluble alkali salts arelobtaiif" "As it is well knowntofth f different acids of the general'tlormula, 0H,

a yea wherein zero or a wholelpositiileifieiand the ring skeleton may be saturated'or 'may contain double bonds. If double-bonds are'present,

5 sessthe general formula: 7

there may be more double bonds hqwerer not more than two double bonds in the same ring. The side chain clinging "to theCl atom lfl fmay bestraight or arborescent V j fIfhe ring skeletonmay, according to the starting material and its treatment, contain various substituents, preferably oxygen containing sub,- stituents, such as hydroxy groupsfesterifledhy droxy groups, ketonic oxygen and the like, igvhicli especially adhere to the Catom 3. i According to the-kind of the sterol subjected to the oxidation there maybe in the oxidation mixture.' acids of the cholesterol series aswell as of the 'dihydr'o-cholsterol' or the coprosterol series; if substituents are present in position 3, the acids in question may 'appert'ainto the normalorto theepi-serie sl Thus; the acids contained m the 5 said ox-idar tion mixtures and therefore'to'be taken into consideration for our process may in the case of dihydro-cholesterol asstarting material be the 3-hydroxy-allocholaic acid,'the 3-hydroxy-bisnorallocholaic acid and the S-hydroxy-aetio-allocholaic acid or, employing the epi-dihydrm cholesterol, the corresponding epi-compounds; starting from coprosterol or epi coprosterol, in the same manner are obtained the' 3-hydroxycholaic acid, the 3-hydroxy-norrcliolaic acid etc., or the epi acids respectively. Ffrom unsaturated s'terols, such "as cholesterolf-stigmasterol; sitojstem; cincholpre'sult unsaturated acids; for in} stance iromcholesterolb v li -hydroxy-choler'iic' acid,

3-hyd'roxyi-nor-cholenic acid, 1 3- hydroxy bisnor-- cholenic acid and "3-hydroxy aetio cholenic acid, i.;' e. ratherthe B aCyIated acids. When no "protection or the hydroxy groups has beeneiiected before the oxidation or when starting from a ketonic derivative of sterols, 'e". "g; cholestenone, one "obtains keto'ni'c-acids; such" as,' for example. 3-'ketof-cholenic acid; j

1 As we have now'found, there are to be'attained primary'alcohols'of physiological activity'by re duction of derivatives of'thse acids, which pos- V CH3? "I If -(OH2),.OOR

wherein n is zero or a whole, positivefigure; and

they are especiallyiin theipositioniiiT612155; \Ri.=a1koxyi.ore-halogen 51, The; esterificatiqn oy and reduces for instance the obtained methyl.

ester by means of sodium and alcohol according to Bouveault and Blanc (see Comptes Renduside lAcadmie des Sciences, tome 136,;p'age' 1676). The mixture of acids may be esterified as awhole tioned methods, a hydrogenation of nuclear dour ble bonds generally does not take place. Sometimes it is not necessary to saponify the acids,

.because by the said methods of reduction a saand the mixture of the resulting esters s'ubse=,.i"'

quently be reduced, wherebya mixturofphys'io-ff logically effective alcohols is obtained, which may a be applied in therapeutics as such or may be separated into its several parts. But one may also first separate the mixture of acids intothe single constituents by various methods and transform the acids each for itself into the respective primary alcohol.

In many cases it'ma'y be preferable 'to' prepare the halides, 'e. g. the chlorides, of the acids instead of the esters, for instanceby treating' the acids with phosphorus chlorides or thion'yl chloride. The obtained chlorides may now be reduced to the primary alcohols catalytical1y, for instance according tothe method of Rosenmund (see -Bericlite der Deutschen Chemischen' Gesellschaf t, 1921, tome 54, page 638).

In this case one may also work up" the mixture of acids direct, i. e. without separatingit into its components, or reduce the single individuals Therefore, the primary alcohols obtainable'ac cording to the above mentioned processes and being, the subject of our application possess the following general formula:

z 4 on, cno onzon :wherein again 11. is zero or a whole, positive figure.

The side chain may again bestraight or arborescent. Especially those alcohols deserve'interest where the CHzOH-group is fixed immediately to the C atom 17 or is separated from it .byone, two or threeCI-Iz-groups; although in r I many cases alcohols with a; longer side chain may be of importance. The ring skeleton may be;saturated or unsaturated, it, may carry subfstituents or may be unsubstituted. Particularly interesting are again the alcohols'which possess a double bond in position 4,5'or 5,6 and carry in position 3 an oxygen containing group, e.;g.- a hydroxy group. Of course, the nature of the obtained alcohol depends to. a high, degree? upon the sterol or sterol derivative subjected to the in positionB one may by acetylating the hydroxy group take care that the acids, which-form the yield.

p'onificationto the hydroxy groups may happen. "I'hus, one obtains the same 3-hydroxy-choleny1- methanol starting either from the 3-acetoxy-cholenic acid methyl ester or the 3-hydroxy-cholenic acid methyl ester. If an originally existing ketonic gram-e. g. in position 3, is desired to be methan'ol, 3ahydroxy aetio cholenylmethanol,- 3'- epi-hydroxy-allocholanyl methanol, 3-epi-hydroxy-aetio-allocholanyl methanol and many others. Y

Of" course, these examples are to be considered as a selection for demonstration only and signify by no means a limitation to the said alcohols.

The alcohols obtained according to our invention possess physiological activity, preferablythe activity of male sexual hormones. Now, the acid constituents of the reaction mixtures attained at'theoxidation'of sterols etc. being consider.- able inquantity and, furthermore, the reactions applied for the transformation of the acid constituents into the primary alcohols being chief reactions which pass with good yield, our invention therefore renders it possible to prepare substances of the'activity of a male sexual hormone, for example, in a simple manner and with good The new alcohols are well crystallisable, colorless compounds, which melt, as a rule, above 100 0., and which are'soluble in the ordinary organic solvents, such as ether, acetone, alcoholspreferably low aliphatic alcohols-and insoluble or difficultly. soluble in water, acids and alkalies.

They possess optic activity and form well crystallisable derivatives, e.g.' diacetates, which are likewise optically active.

' Examples 1) On" oxidation of cholesterol acetate dibromide with chromic acid. and debromination of the oxidation mixture (see for. example Helvetiacid fronr'stigmasterol acetate :dibromide and phytos'terol facetate dibromide *(see for example British patent specification 453,773); On-saponflying-the acetates one, obtains the free acids; which have also been described inliterature,

g'fo'f the acid constituents contained in the oxidation" mixture of cholesterol acetate dibromide and-isolated therefrom -forjinstance accord ing to the methods. applied in the "above mentioned Britishpatent specification are dissolved in 250 cc. of i'nethanoLand 2 cc. of sulphuric acid are added. This solution is heated for one hour with reflux.- After coolin'g'the mixture of .esters isprecipitated by addingwater and recrystallised from acetone. v

The carefully dried mixture of esters. which isobtaified in a yield of about 10 grams is now dissolved in 3-4 times the quantity of absolute alcohol. Thissolution is droppedonto 5. grams of coarsely cut sodium. After heating several hours under reflux'the whole of the sodium is dissolved by'addlng sufficient absolute alcohol, whereupon heating on the steam-bath withrefiux is continued'for another. hour. Now thesolution is still more diluted with water and repeatedly shaken outwith ether. The unitedethereal extracts are washed neutral, dried and evaporated. The residue represents a mixture of primary alcohols of which the following have been iisolated; H ,v

3-hydroxy aetio cholenyl methanol, meltingpoiht" 196 199, CzoHazO'z: talc. 78.93% C, 10.53% H, found 79.02% C, 10.59% H.

3-hyclroxy-bisnor cholenyl methanoL-meltingk I H;'foun d 79.3% c;10.s% H. v 3- hydroxy -'ch'olenyl methanol, melting-point 196-1972 024114002: calc.' 80.0 c, 11.1% H, found 79.5% C, 1 1.1% H; 1-

[a]p=}-33' (in pure ethyl-alcohol) (2) Epi-dihydro cholesterol is oxidised with chromic acid as it is described for examplein Helvetica Chimica' Acta, tome XVII, page 1403:

A solution'of g. of chromium trioxidein 30 4 cc. of Water and1150 cc. of'glacial acetic acid is added drop by drop to 20 g. of epi-dihydro-choe lesterol acetate in'about one litre of glacial acetic acid while stirring and heating. After heating several further hours surplus chromium'trioxide is..destroyed by. addition of methanoL; Thereupon the glacial acetic acid isdistilled oil in vacuo; the residue is taken up with water and repeatedly shaken with ether. The united ethereal .extracts are thoroughly washed with water and thereupon shaken with about 10% sodium hydroxide lye in order to separate the ac d Darts, whereby diiiicultly soluble-sodium salts precipitate. By decomposing these salts withdilute sulphuric acid one obtains a mixture of acetates of acids of the allocholane series which differ in the number of C atoms contained in the side chain. By means of usual methods one may separate the mixture of acids into the single components, thus obtaining the following acids: 3-epi-acetoxy allocholaic acid, 3-epi-acetoxy-nor-allocholaic acid, 3-epiacetoxwb'isnor allocholaic;; acid, ,3=epi-acetoxy.- aetidsallocholaic acidi These acids; lesp fiially .the not este'rified acids; are" described for examplein Berichte .'der*;- Deutschen Qhernischen- Qesell;

J The:single acids are-suspended in 2 0ti1nesthe quantity'of etherg and an excess of ane in ethereal solution is-added. =:Afte standing several hours the acid is dissolved. whereby this sapqnified- "Thus; he j. pr mar 1 process may be a'cceleratedby' adding some'ymeth anol. The surplus diam-methane is destroyed by some cubiccentimetres of acetic acid-,angl the yolatile parts of thereactioninixture are distilled 01f. The residue represents themethyl' ester of the employed acidj which may be subjected without further puriflcation'to the reduction as it is described in Example Qne obtains thus the iollowing primary 8.1CO1101S:" (a) 3-epi-hydroxy-allocholanyl methanol, -melt-. ing-point 187,- 189? --C-., [-ln=+36 (in abs. 7 alcohol), calc. 79.49 c, 11.59% rou d 79.78% C,-1'1;.77%H. U l The compound-*forms a diacetate which melts at 1'14 116-; ,[mln='t34 (in abs. al-

cohol). cams-75.28% -C,- 10.31% H; found 15.12%0, 19.43%11. (11) -3l-epi-hydroxy nor allocholanyl methanol, melting point 213-21 3.5-C., [a]l)=+31 (in ;abs; alcohol)','calc. 79.24% C, 11.48% H; 1 found 79.15%-C,'11 .34% H Diacetate: melting-point 135-137 (3.. [1111): "345 ':(ir 1 'abs.-- alcohol), calc. 74.94% C, 10.18%H, found. 75.03% C, 10.01% H. (0) 3 epi-hydr0xy+bisnor allocholanyl methanol, me1ting-point' 230 232 Cs; [a]D'= 27 '(in 1 abs. alcohol).,=calc:'78.97% C, 11.37% H, -1found79.32%.C,;11.51% H.1 1

. 3 Diacetate: melting point146-14750.[01113:

10.04% H, found 7416071 C 10.21% H.

(d)v 3-epi-hydr yw tiq-allocholan lz: m th n melting-point122l-223 .-,';[{]p t22 (in 8 i)? Gi -10% :zH", .j 1nd' 8.09.%" 0.11.. i. Diacetater melting-mint;-1Q6'C; [n]: +16 (inf-abs.alcohol),;.icalb.::f 3.83%1C. I 9.74%"H,iound 73.91%YC;-9.87.%1 f Instead of. separatingthe mixtur o f acids into the single acids one may' a lso esterifyas a whole, wherebya mixture of saidj al'cohols is. Obtained. This mixture may. be applied directin' therapeutics; thus saving; the various procedures of sepsiration. '1.

(By-The mixture of acids obtained on-oxidising epi-dihydrocholesterol (see Example 12) is''i nstead of'esterifyingihtransformed to the" corresponding mixtureo acidchloridesbymeans of thionyl chloride." Y

' ,5 grams of this mixture are, ,d isso1ved in 100 cc, of X'Y1Q1. 'I'o this mixturejz ii drops of; chin olin andv 3 grams bariumj sulphate-palladium cataly} ser; are added. Qne nyqrq nisees hours' 'at a bath temperature of 120 1f3 0 f- C 'Ijhe reupo n the catalyser is filtered off and the x910] layer shaken nss stite wit d lu :as ii a After distilling r the xyl'ol n; vacuo we ob- :ta nsjat rema n. a ua i y f o 'rams. whichj is .trahsform'edtd aci esters by heating 1 W .eht 'an y ride "The estersare sepaiatd r m the re o xt re on th is 'e th ii he ris lubiIiw: in alkalis and; subsequ gy $01 9 5. d r 4. 5 M 9b.-

sprm c; in; Exam le 2' de 9 mi ed-@171" 2 Various changes may heima dein thedetails disclosed in the f regoing speci fioajti'olflwitholit departing fromithe invention .or sac ficlng aduantagesthereof. In the claims atlixed w this speci c tion m5 selection of d any particular modification 'of f the evcnt o is. n'te to ,th clx ib i ig-ctmr modifications thereof, and the 7 right tosubsequently, make claim toany modification not covered by these claims'is expressly reserved. v

Weclaimz n 1. Process of preparing primary alcohols with physiological activity consisting in separating the carboxylic acids,.which arise onoxidation of compounds; with the cyclopentan'o. h'ydrophen anthrene skeleton and aside chain, in position 17 of this skeleton. from-the;m1-xture off jthe oxide tion products by sapon'ifying' their'acetyl comby saponifying theiracetyl compounds and transforming their esters tofprimaryalcohols by reduction.

3. Process of preparing primary alcohols with physiological activity oxygen containingcompounds withthe cyclopentano hydrophenanthreneskeleton and a side chain in position 17 fof 'this skeleton, from the mixture of the oxidation products by saponifying consisting in separating the carboxylic acids,which arise on oxidation of dation products by saponifying their acetyl compounds, transforming them to, halides". with the, carboxylic acids, which garisefon' oxidation of their acetylcompounds and. transforming their esters to primary alcohols by'reduction;

4. Process of preparing primary alcohols with physiological 'acti v ity 1 consisting in separating the carboxylic acids, whicharise on oxidation of 3-hydroxy-A5,6-compounds with the cyclopentano hydrophenanthrene skeleton and a side chainin position 17 of this skeleton, from the mixture of the oxidation products by saponifying their. acetyl compounds and transforming their esters toprimary alcohols by reduction. j 5. Process of preparing primary alcohols with physiological activity consisting in separating the carboxylic acids, which arise on oxidation of 3-acyloxy-A5,6-compounds with the CYCIOPBH! tano hydrophenanthrene skeleton and a' side chain-in position 17 of this skeleton, fromthe mixture of the oxidation products bysaponifying their acetyl compounds and transforming their esters to primary alcohols by reduction. Y 6. Process of preparing primary alcohols with physiological activity consisting in separating the carboxylic acids, which arise on oxidation of 3-keto- A5,6-compounds with the; cyclopentano hydrophenanthrene skeleton and a side chain'in position 17 of this skeleton, from the mixture of the oxidation products by 'saponifyingjf their acetyl compounds and transforming their esters to primary alcoholsby reduction.

7. Process of preparing primary alcohols with physiological activity consisting in separating the carboxylic acids, which ariseon-oxidation of compounds with the cyclopentano hydrophene anthrene skeleton and a side chain inlposition 17 of this skeleton, from the mixture of the oxidation products by saponifying their acetyl com pound, transforming them to esters, precipi tating the esters with water and reducing'the latter to primary alcohols.

8. Process of preparing primaryalcohols with physiological activity consisting in' separatin the carboxylic acids, which arisefon oxidation of compounds with the cyclopenta'no' hydrophenanthrene skeleton and a side .,chain.in position 17 of this skeleton,-from the of the oxi tano, 'hydrophenanthre ne skeleton and a side chain ,in position 17 pflth kel'etom from the 'r'nixturelof theoxid tion'pro'clu'cts bysaponifying their jacetyl compound, jtransformingythem to esters, precipitating the"festers with water and reducing the-"latter to primary 'alcoh'ols: f l 10. Process of preparing primary alcohols with physiological activityconsisting inseparating the carboxylicacids', whichlarise on oxidation of 3- acyloxy-'A5',6-,compounds.- with the. 'cyclopentano hydrophenanthrene', skeleton and a side 'chain in position".17"of ,t'his skeleton," from. the mixture: of the oxidation products by saponifying their acetyl compound, transforming themftotestersand re ducing the latter to primary alcohols.

l1. Processof preparing primary alcohols with physiological activity consisting in'separating the carboxylic acids,' which arise on oxidation oraacyloxy-ni'fi 'compounds with the 'cyclopentano hydrophenanthrene skeleton and a side chain in position 1!? of this skeleton,-from the mixture of the oxidation products by saponifying their acetyl compound, transforming them to esters, reducing the esters to primary alcohols and saponifying thelatter.

12. Process of preparing primary alcohols with physiological activity consistingjn separating the carboxylic acids, which arise-on oxidation of 3- acy1oxy-A5,6comp0unds with the cyclopentano hydrophenanthrene skeleton anclva side chain in position 17 of this skeleton, from the mixture of the oxidation. products, saponifyingthem, trans,- forming the free acids to esters, precipitating the esters with water and reducing, the latter to primary alcohols. 13'. Processor? preparing primary alcohols with physiological activity consisting in separating the carboxylic acids, which .arise on oxidation of 3- keto-A5,6-compou'nds with the cyclope'ntano hydrophenanthrene skeleton and aside chain in position '17 of this skeleton, from :the mixture of the oxidation products by saponifying their acetyl compound, transforming them to .esters,

precipitating the esters with water and reducing the latter to primary alcohols. r s

l4 Unsaturated primary alcohols of the gerieral formula: I f f 2)n-CH2?H inwater, alkalis, t

15. Unsaturated primary alcohols of the general formula:

CH3 CH:

a l /\V wherein n is zero or a whole positive figure and the cyclopentano hydrophenanthrene skeleton contains a double bond in position 5,6, which are colorless, crystalline, optically active, physiologically effective, soluble in alcohols, ether, acetone, and diflicultly soluble in water, alkalis, acids.

16. Unsaturated primary alcohols of the general formula:

wherein n is zero or a whole positive figure and the cyclopentano hydrophenanthrene skeleton contains not more than two double bonds in the same ring, one of these double bonds being in 4,5 or 5,6 position and an oxygen group in position 3, which are colorless, crystalline, optically active, physiologically effective, soluble in alcohols, ether, acetone, and diflicultly soluble in water, alkalis, acids.

18. Unsaturated primary alcohols of the general formula:

CH3 CH3 wherein n is zero or a whole positive figure and.

19. Unsaturated primary alcohols of the general formula:

wherein n is zero or a whole positive figure and the cyclopentano hydrophenanthrene skeleton contains an oxygen group in position 3 and a double bond in position 5,6, which are colorless, crystalline, optically active, physiologically effective, soluble in alcohols, ether, acetone, and difflcultlysoluble in water, alkalis, acids.

20. The primary alcohol of the constitution of a 3-hydroxy-A5,6-cholenyl methanol, which is colorless, crystalline, optically active, physiologically efiective, soluble in alcohols, ether, acetone, difiicultly soluble in water, alkalis, acids, and melts at 196-197.

21. The primary alcohol of the constitution of a 3-hydroxy-A5,6 bisnor cholenyl methanol,

which is colorless, crystalline, optically activephysiologically effective, soluble in alcohols, ether, acetone, diflicultly soluble in water, alkalis, acids, and melts at 202206.

22. The primary alcohol of the constitution of a 3-hydroxy-A5,6-aetio-cholenyl methanol, which is colorless, crystalline, optically active, physiologically efiective, soluble in alcohols, ether, acetone, difiicultly soluble in water, alkalis, acids and melts at 196-199".

23. Unsaturated alcohols of the general formula:

CH3 CH3 wherein n is an integer including zero and where in X is selected from the group consisting of hydrogen and hydroxyl and the cyclopentanohydrophenanthrene ring contains not more than two double bonds in the same ring, one of these double bonds being in 4,5 or 5,6 position, which are colorless, crystalline, optically active, physiologically efiective, soluble in alcohols, ethers, acetone, and diflicultly soluble in water, alkali, and acids.

FRITZ JOHANNESSOHN. HANS HATZIG. 

